Design and quantitative evaluation of 'Aerosol Bio-Containment Device (ABCD)' for reducing aerosol exposure during infectious aerosol-generating events.

The Coronavirus Disease 2019 (COVID-19) pandemic renewed interest in infectious aerosols and reducing risk of airborne respiratory pathogen transmission, prompting development of devices to protect healthcare workers during airway procedures. However, there are no standard methods for assessing the efficacy of particle containment with these protective devices. We designed and built an aerosol bio-containment device (ABCD) to contain and remove aerosol via an external suction system and tested the aerosol containment of the device in an environmental chamber using a novel, quantitative assessment method. The ABCD exhibited a strong ability to control aerosol exposure in experimental and computational fluid dynamic (CFD) simulated scenarios with appropriate suction use and maintenance of device seals. Using a log-risk-reduction framework, we assessed device containment efficacy and showed that, when combined with other protective equipment, the ABCD can significantly reduce airborne clinical exposure. We propose this type of quantitative analysis serves as a basis for rating efficacy of aerosol protective enclosures.

Fluid dynamic simulations at the interface of the blue-green sharpshooter functional foregut and grapevine xylem sap with implications for transmission of Xylella fastidiosa.

Xylella fastidiosa is a multi-continental, lethal, plant pathogenic bacterium that is transmitted by sharpshooter leafhoppers (Insecta: Hemiptera: Cicadellidae: Cicadellinae) and adult spittlebugs (Hemiptera: Aphrophoridae). The bacterium forms biofilms in plant xylem and the functional foregut of the insect. These biofilms serve as sources of inoculum for insect acquisition and subsequent inoculation to a healthy plant. In this study, 3D fluid dynamic simulations were performed for bidirectional cibarial propulsion of xylem sap through tube-like grapevine xylem and an anatomically accurate model of the functional foregut of the blue-green sharpshooter, Graphocephala atropunctata. The analysis supports a model of how fluid dynamics influence X. fastidiosa transmission. The model supports the hypothesis that X. fastidiosa inoculation is mostly driven by detachment of bacteria from the foregut due to high-velocity flow during egestion (outward fluid flow from the stylets). Acquisition occurs by fluid dynamics during both egestion and ingestion (fluid uptake through the stylets and swallowing). These simulation results are supported by previously reported X. fastidiosa colonization patterns in the functional foregut and sharpshooter stylet probing behaviors. The model indicates that xylem vessel diameter influences drag forces imposed on xylem wall-adherent bacteria; thus, vessel diameter may be an important component of the complex transmission process. Results from this study are directly applicable to development of novel grapevine resistance traits via electropenetrographic monitoring of vector acquisition and inoculation behaviors.

Functional foregut anatomy of the blue-green sharpshooter illustrated using a 3D model.

Sharpshooter leafhoppers (Hemiptera: Cicadellidae: Cicadellinae) are important vectors of the plant pathogenic bacterium Xylella fastidiosa Wells et al. (Xanthomonadales: Xanthomonadaceae). This pathogen causes economically significant diseases in olive, citrus, and grapes on multiple continents. Bacterial acquisition and inoculation mechanisms are linked to X. fastidiosa biofilm formation and fluid dynamics in the functional foregut of sharpshooters, which together result in egestion (expulsion) of fluids likely carrying bacteria. One key X. fastidiosa vector is the blue-green sharpshooter, Graphocephala atropunctata (Signoret, 1854). Herein, a 3D model of the blue-green sharpshooter functional foregut is derived from a meta-analysis of published microscopy images. The model is used to illustrate preexisting and newly defined anatomical terminology that is relevant for investigating fluid dynamics in the functional foregut of sharpshooters. The vivid 3D illustrations herein and supplementary interactive 3D figures are suitable resources for multidisciplinary researchers who may be unfamiliar with insect anatomy. The 3D model can also be used in future fluid dynamic simulations to better understand acquisition, retention, and inoculation of X. fastidiosa. Improved understanding of these processes could lead to new targets for preventing diseases caused by X. fastidiosa.

Escherichia coli O157:H7 and Salmonella Typhimurium adhesion to spinach leaf surfaces: Sensitivity to water chemistry and nutrient availability.

This study investigated the effects of solution chemistry and growth conditions on bacterial deposition on spinach leaf surfaces using a parallel plate flow cell. Two food safety pathogens of concern and two non-pathogen bacterial surrogates (environmental E. coli isolates) were grown in ideal (LB media) and nutrient-restricted (M9 media) conditions. Bacterial attachment was quantified as mass transfer rate coefficients for cells suspended in 10 mM KCl, CaCl2 and artificial groundwater, and cell and leaf surfaces were extensively characterized (zeta potential, hydrophobicity, extracellular polymer (EPS) composition). Between the pathogens, E. coli O157:H7 attachment was greater than that of Salmonella Typhimurium, attributed to measurable variability in cell surface charge and hydrophobicity. When grown in M9 media, both pathogens were significantly more adhesive to spinach surfaces (p < 0.01) than when grown in LB media. Surrogates did not follow this trend and showed minimal changes in adhesion kinetics and surface properties between growth conditions. EPS sugar/protein ratios were reduced in some of the highest attachment scenarios, suggesting that changes in EPS composition in favor of proteins may play a role. These results show the importance of growth conditions and solution complexities in understanding mechanisms of aqueous bacterial adhesion to food surfaces.

Influence of septic system wastewater treatment on titanium dioxide nanoparticle subsurface transport mechanisms.

Engineered nanomaterials (ENMs) are commonly incorporated into food and consumer applications to enhance a specific product aspect (i.e., optical properties). Life cycle analyses revealed ENMs can be released from products during usage and reach wastewater treatment plants (WWTPs), with titanium dioxide (TiO2) accounting for a large fraction. As such, food grade (FG) TiO2, a more common form of TiO2 in wastewater, was used in this study. Nanomaterials in WWTPs have been well characterized, although the problematic septic system has been neglected. Elution and bioaccumulation of TiO2 ENMs from WTTPs in downriver sediments and microorganisms has been observed; however, little is known about mechanisms governing the elution of FG TiO2 from the septic drainage system. This study characterized the transport behavior and mechanisms of FG TiO2 particles in porous media conditions after septic waste treatment. FG and industrial grade (IG) TiO2 (more commonly studied) were introduced to septic tank effluent and low-ionic strength electrolyte solutions prior to column transport experiments. Results indicate that FG TiO2 aggregate size (200-400 nm) remained consistent across solutions. Additionally, elution of FG and IG TiO2 was greatest in septic effluent at the higher nanoparticle concentration (100 ppm). FG TiO2 was well retained at the low (2 ppm) concentration in septic effluent, suggesting that particles that escape the septic system may still be retained in drainage field before reaching the groundwater system, although eluted particles are highly stabilized. Findings provide valuable insight into the significance of the solution environment at mediating differences observed between uniquely engineered nanomaterials. Graphical abstract.

Comparison of filtration mechanisms of food and industrial grade TiO2 nanoparticles.

The removal of food and industrial grade titanium dioxide (TiO2) particles through drinking water filtration was assessed via direct visualization of an in situ 2-D micromodel. The goal of this research was to determine whether variances in surface composition, aggregate size, and ionic strength result in different transport and deposition processes in porous media. Food and industrial grade TiO2 particles were characterized by measuring their hydrodynamic diameter, zeta potential, and zero point of charge before introduction into the 2-D micromodel. The removal efficiency as a function of position on the collector surface was calculated from direct visualization measurements. Notably, food grade TiO2 had a lower removal efficiency when compared with industrial grade. The difference in removal efficiency between the two particle types could be attributed to the higher stability (as indicated by the larger zeta potential values) of the food grade particles, which lead to a reduced aggregate size when compared to the industrial grade particles. This removal efficiency trend was most pronounced in the rear stagnation point, due to the high contribution of hydrodynamic forces at that point. It could be inferred from the results presented herein that particle removal strategies should be based on particle aggregate size and surface charge. Graphical abstract ᅟ.

Linking microbial community structure to function in representative simulated systems.

Pathogenic bacteria are generally studied as a single strain under ideal growing conditions, although these conditions are not the norm in the environments in which pathogens typically proliferate. In this investigation, a representative microbial community along with Escherichia coli O157:H7, a model pathogen, was studied in three environments in which such a pathogen could be found: a human colon, a septic tank, and groundwater. Each of these systems was built in the lab in order to retain the physical/chemical and microbial complexity of the environments while maintaining control of the feed into the models. The microbial community in the colon was found to have a high percentage of bacteriodetes and firmicutes, while the septic tank and groundwater systems were composed mostly of proteobacteria. The introduction of E. coli O157:H7 into the simulated systems elicited a shift in the structures and phenotypic cell characteristics of the microbial communities. The fate and transport of the microbial community with E. coli O157:H7 were found to be significantly different from those of E. coli O157:H7 studied as a single isolate, suggesting that the behavior of the organism in the environment was different from that previously conceived. The findings in this study clearly suggest that to gain insight into the fate of pathogens, cells should be grown and analyzed under conditions simulating those of the environment in which the pathogens are present.

Pseudomonas aeruginosa attachment on QCM-D sensors: the role of cell and surface hydrophobicities.

While biofilms are ubiquitous in nature, the mechanism by which they form is still poorly understood. This study investigated the process by which bacteria deposit and, shortly after, attach irreversibly to surfaces by reorienting to create a stronger interaction, which leads to biofilm formation. A model for attachment of Pseudomonas aeruginosa was developed using a quartz crystal microbalance with dissipation monitoring (QCM-D) technology, along with a fluorescent microscope and camera to monitor kinetics of adherence of the cells over time. In this model, the interaction differs depending on the force that dominates between the viscous, inertial, and elastic loads. P. aeruginosa, grown to the midexponential growth phase (hydrophilic) and stationary phase (hydrophobic) and two different surfaces, silica (SiO(2)) and polyvinylidene fluoride (PVDF), which are hydrophilic and hydrophobic, respectively, were used to test the model. The bacteria deposited on both of the sensor surfaces, though on the silica surface the cells reached a steady state where there was no net increase in deposition of bacteria, while the quantity of cells depositing on the PVDF surface continued to increase until the end of the experiments. The change in frequency and dissipation per cell were both positive for each overtone (n), except when the cells and surface are both hydrophilic. In the model three factors, specifically, viscous, inertial, and elastic loads, contribute to the change in frequency and dissipation at each overtone when a cell deposits on a sensor. On the basis of the model, hydrophobic cells were shown to form an elastic connection to either surface, with an increase of elasticity at higher overtones. At lower overtones, hydrophilic cells depositing on the hydrophobic surface were shown to also be elastic, but as the overtone increases the connection between the cells and sensor becomes more viscoelastic. In the case of hydrophilic cells interacting with the hydrophilic surface, the connection is viscous at each overtone measured. It could be inferred that the transformation of the viscoelasticity of the cell-surface connection is due to changes in the orientation of the cells to the surface, which allow the bacteria to attach irreversibly and begin biofilm formation.

Dynamics of oscillatory phenotypes in Saccharomyces cerevisiae reveal a network of genome-wide transcriptional oscillators.

Genetic and environmental factors are well-studied influences on phenotype; however, time is a variable that is rarely considered when studying changes in cellular phenotype. Time-resolved microarray data revealed genome-wide transcriptional oscillation in a yeast continuous culture system with ∼ 2 and ∼ 4 h periods. We mapped the global patterns of transcriptional oscillations into a 3D map to represent different cellular phenotypes of redox cycles. This map shows the dynamic nature of gene expression in that transcripts are ordered and coupled to each other through time and concentration space. Although cells differed in oscillation periods, transcripts involved in certain processes were conserved in a deterministic way. When oscillation period lengthened, the peak to trough ratio of transcripts increased and the fraction of cells in the unbudded (G0/G1) phase of the cell division cycle increased. Decreasing the glucose level in the culture medium was one way to increase the redox cycle, possibly from changes in metabolic flux. The period may be responding to lower glucose levels by increasing the fraction of cells in G1 and reducing S-phase gating so that cells can spend more time in catabolic processes. Our results support that gene transcripts are coordinated with metabolic functions and the cell division cycle.

Impact of growth conditions on transport behavior of E. coli.

The aim of this investigation is to determine the effect that growth solution has on the cell surface properties and transport behavior of eleven Escherichia coli isolates through saturated porous media. The two growth solutions used were a standard laboratory growth medium (LB) and a dairy manure extract solution. In general, cells grown in manure extract were more hydrophobic, had a more negative zeta potential, had lower amounts of surface macromolecules, and had lower attachment efficiencies than isolates grown in LB. An inverse relationship between the natural log of zeta potential and the attachment efficiency of the isolates for the cells grown in LB media was the only statistically significant correlation observed between transport behavior and cell characteristics of the isolates. This study shows the need to consider growth conditions when studying bacteria to better mimic the environmental stresses that bacteria undergo in the natural environment. This approach could lead to a better understanding of the behavior of manure-derived bacteria in aquatic and terrestrial environments.

Correlating transport behavior with cell properties for eight porcine Escherichia coli isolates.

In this study we investigate how growth stage and depositional environment affect variability of cell properties and transport behavior of eight porcine E. coli isolates. We compared the surface properties for cells harvested during exponential and stationary growth phase and their transport behavior through columns packed with either uncoated or Fe-coated quartz sand. We then investigated correlations between measured cell properties and fitted bacterial attachment efficiencies. For both growth stages we found that bacterial attachment efficiencies in the uncoated quartz sand varied among the eight different isolates by over an order of magnitude whereas attachment efficiencies in the Fe-coated sands varied by a factor of less than two. With the exception of one isolate, growth condition had minimal impact on attachment efficiencies to the uncoated sands. A strong and statistically significant inverse relationship was observed between bacterial attachment efficiencies in the uncoated quartz sand columns and log-transformed zeta potential, whereas a mild yet statistically significant relationship between bacterial attachment efficiencies in the Fe-coated sands and cell width was observed. For the experimental conditions used in our study, we found that variability in E. coli transport was more dependent on the depositional environment than on growth conditions.